Enhanced glucocorticoid exposure facilitates the expression of genes involved in prostaglandin and estrogen syntheses in bovine placentomes at induced parturition.

The mechanism by which the fetal membrane detaches after parturition in cattle is poorly understood, but the upregulation of placentomal prostaglandin and estrogen synthesis are considered to be important. This study investigated whether enhanced glucocorticoid exposure affected the functional maturation of placentomes at induced parturition. Placentomes were collected immediately after spontaneous (beef; n = 5, dairy; n = 5) or induced parturition in beef and dairy cattle. Parturition was induced conventionally using prostaglandin F2α (beef; n = 7, dairy; n = 6) or dexamethasone (beef; n = 6) or with a combination of triamcinolone acetonide (a long-acting glucocorticoid) and a high dose of betamethasone (TABET treatment, beef; n = 6, dairy; n = 9). Gene expression levels and protein localization in placentomes were analyzed by RT-qPCR and immunohistochemistry, respectively. Compared with the conventional methods, TABET treatment resulted in upregulated PTGS2 expression in cotyledons. The expression levels of PTGS2 and PGES were positively correlated in both cotyledons and caruncles. TABET treatment also upregulated the expression of CYP17A1, but not of CYP19A1, in cotyledons. The results revealed, for the first time, that PLA2G4A was localized in microvascular endothelial cells in the cotyledonary villi and the maternal septum. PTGS2 and PGES were colocalized in mononucleated cells of the cotyledonary villi and caruncle epithelial cells adjacent to the chorionic plate. TABET treatment upregulated the expression of placentomal genes involved in PGE2 synthesis and the conversion of pregnenolone to androstenedione. Thus, enhanced glucocorticoid exposure might partially facilitate the functional maturation of placentomes at induced parturition in cattle.

A novel stereotaxic approach to the hypothalamus for the use of push-pull perfusion cannula in Holstein calves.

To determine secretory patterns of growth hormone-releasing hormone (GHRH) and somatostatin (SS) and their roles in the regulation of growth hormone (GH) secretion, a method for collecting hypothalamic perfusates, a push-pull perfusion method was developed in calves. With the use of the stereotaxic apparatus for cattle, a cannula was implanted into the hypothalamus of four male calves based upon cerebral ventriculography. Push-pull perfusates were collected at 10 min intervals for 6h and GHRH and SS concentrations in perfusates and plasma GH concentration were determined by EIAs and RIA, respectively. A cannula was implanted into the hypothalamus based on the image of the third ventricle and maintained for 1 month. GHRH and SS showed pulsatile secretion and the pulses for GHRH and SS were irregular in conscious animals. Neither GHRH nor SS secretion had a clear relationship with GH secretion. In the present study, we thus (1) established a stereotaxic technique for approaching the hypothalamus using cerebral ventriculography for calves, and (2) demonstrated that GHRH and SS secretion were pulsatile but not closely related to GH profile in conscious calves. The technique is useful for the study of the functions of the hypothalamus in the control of pituitary hormones in cattle.

A technique for long-term implantation of a microcatheter into the third ventricle of post-pubertal Chinese Meishan pigs based on ventriculography.

The objectives of this study were to implant a microcatheter into the third ventricle of post-pubertal Chinese Meishan pigs, and to maintain the microcatheter for a long time without causing stress. Fourteen pigs (45-60 kg BW) were used. Each pig was anesthetized and the head was orientated on the stereotaxic apparatus. A radiopaque dye was placed into the ventricle via a guide cannula inserted 3.5 mm forward of the bregma. A microcatheter was inserted into the third ventricle using ventriculography, and fixed with dental cement to a metal-mesh protector and screw anchors embedded into the skull. The opposite end of the microcatheter was externalized from the dorsal neck so that corticotropin-releasing hormone (CRH) could be injected easily. Simultaneously, a catheter was fitted in the jugular vein, and the free end of the catheter was externalized from the dorsal neck. Microcatheter-implanted pigs showed a normal progesterone concentration profile, and a constant cortisol level during at least two estrous cycles. Furthermore, intracerebroventricular injections of CRH (25 microg/500 microl) resulted in an increased plasma cortisol concentration (P < 0.05). Thus, the technique developed in this study enables us to approach the third ventricle in post-pubertal freely-moving pigs effectively over a long time, without causing stress.